Microfluidic device utilizing magnetohydrodynamics and method for fabrication thereof

ABSTRACT

Microfluidic channels utilizing magnetohydrodynamics are used to pump very small volumes of solution. The channels have electrodes along the walls and a current carrying species within a solution carries current through the solution. The combination of the electric and magnetic fields causes the solution to flow through the channel.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a divisional of U.S. patent application Ser. No.10/317,777, filed Dec. 22, 2002 and claims priority to U.S. patentapplication Ser. No. 10/026,748, now U.S. Pat. No. 6,733,244, filed Dec.19, 2001 and claims priority to U.S. Provisional Application Ser. No.60/257,331, filed Dec. 20, 2000 and claims priority to U.S. ProvisionalApplication Ser. No. 60/278,275, filed Mar. 22, 2001 and claims priorityto U.S. patent application Ser. No. 10/252,342, filed Sep. 23, 2002.

BACKGROUND OF THE INVENTION

1. Field of the Invention

The present invention relates to microfluidic devices. In particular,the present invention relates to the use of magnetohydrodynamics topropel or mix fluids within microfluidic structures.

2. Prior Art

The field of microfluidics is growing rapidly. There is a strong desireto miniaturize chemical assays. A number of various technologies arecurrently being developed in an effort to develop what has become knownas lab-on-a-chip (LOC) technology. It is believed that thesetechnologies will lead to mobile, small scale chemical testing devices.Such devices would have a variety of applications. Emergency MedicalTechnicians and military medics could use such devices to rapidlyanalyze a person's blood chemistry. Forensic scientists could performDNA analysis at a crime scene, instead of waiting hours or days forresults from a laboratory. Realizing the great potential of suchdevices, there have been many attempts to find a low power method ofaccurately propelling extremely small liquid samples throughmicrofluidic structures. The desired width of these channels is lessthan 1 mm, preferably 500 micrometers or less, preferably 100micrometers or less.

Some of the characteristics sought in a microfluidic propulsion systeminclude high fluid flow rates, the ability to change direction of theflow, minimal power requirements and the ability to effectively propel awide variety of fluids through structures composed of a wide variety ofsolid materials. High fluid flow becomes more difficult as microfluidicstructures become smaller. This is due to increased drag created bymoving along the walls of the microfluidic structure. A small powerrequirement is desired so that devices may be compact and portable.Different microfluidic technologies have advantages and disadvantages inthese areas.

It has been found that by forming a gradient of a hydrophobic filmacross a glass or silica plate, water droplets can be induced to travelalong the gradient. However, this method has only achieved relativelyslow flow rates. In addition it is difficult to scale down to themicrofluidic level of less than 500 micrometers. Hydrophobic films tendto work best on relatively large water droplets. It is impossible tochange flow direction and is only effective on aqueous solutions.

There has been some experimentation in using temperature gradients topropel water through small channels. Although flow is reversible, flowrate is very slow. This technique also requires a relatively large powersupply.

Electrokinetics has been a popular field of study in microfluidics. Itprovides for easy change of flow direction and is suitable for verysmall channels. It is also well suited for separating chemicals.However, electrokinetics suffers from disadvantages. It is verysensitive to the chemical properties of both the fluid being manipulatedand the walls of the channel. In addition, this technology requires highvoltage and can only achieve relatively slow flow rates. Electrokineticsalso will not work in the presence of air bubbles, which are common inmicrofluidic systems. Another disadvantage is that electrokinetics isineffective on organic fluids. Like hydrophobic films, this method onlyworks well on aqueous solutions. Application of a strong current mayalso alter chemicals present in the solution, thereby decreasing theaccuracy of any analyses.

Mechanical methods of pumping fluids through microstructures also poseseveral problems. The mechanical methods usually require valves whichcan complicate fabrication and become clogged. Complex mechanicaldevices, including many valves, are difficult to scale down to smallsizes. In addition, mechanical pumping usually requires a pulsating flowand it does not conveniently allow changes in flow direction.

Centrifugation is inexpensive and adaptable to a wide range of channelsizes. However, the flow direction cannot be reversed and this processusually involves a single-use cartridge. Centrifugation also requires alarge power supply. These power requirements rapidly increase and themicrofluidic structure size decreases due to drag.

There is a need for alternative non-mechanical pumping systems that arelower power, operate with a wider range of device materials andsolutions compositions, offer multi-use capabilities, and allow easychange in flow direction.

Magnetohydrodynamics (MHD) has been proposed as an alternative methodfor microfluidic propulsion. This technology involves the application ofa magnetic field and a electric field. The two fields are appliedperpendicular to each other and perpendicular to the desired directionof flow. These fields induce fluid flow perpendicular to both fields.This is known as a Lorentz force. On larger scales, the Lorentz force istoo weak for any practical applications and until recently has beenconsidered only a curious phenomenon.

MHD works best when current density is high, and most electrodes havefairly low current density. However, because of the physics unique tosmall scale diffusion, microelectrodes exhibit very high currentdensities. MHD is therefore much more practical at very small scales.Relatively little power, less than one volt, can achieve high flow ratesin microfluidic structures.

MHD is very susceptible to change of flow direction. By simplyalternating the electrodes, the direction of fluid flow reverses.Similarly, reversing the magnetic field will also reverse flowdirection. The ease of change in flow direction coupled with low powerand high flow rate make MHD an excellent mechanism for microfluidicpropulsion. In addition, Lorentz forces apply to all fluids, so that MHDmay effectively propel both aqueous and organic solutions. MHD is alsounaffected by the materials used to construct microfluidic structures.

There have been limited attempts to apply MHD technology tomicrofluidics already. It has been used successfully on molten metalsand mercury. However, these generally involve high temperatures and arenot well suited to be used in conjunction with chemical assays. Thesemethods have high power requirements and chemical assays are generallynot designed to utilize molten metals.

More recently, attempts have been made to apply MHD to aqueoussolutions. Channels have been constructed having electrodes on opposingwalls. A magnetic field is then applied perpendicular to both thedirection of flow and the electric field generated by the electrodes.Unfortunately, a significant problem has arisen due to waterelectrolysis. Although insignificant on larger scales, bubbles formed bywater electrolysis within a microstructure pose serious problems. Asidefrom blocking fluid flow, they also disrupt the electric field. This inturn disrupts the Lorentz forces and halts fluid flow completely. Onlyvery low voltage, which results in very slow flow rates, have been shownto be practical. At higher voltages, water electrolysis makes MHDimpossible. In addition, MHD is ineffective when applied to hydrophobic,oily solutions that have dielectric points greater than that of water.

There have been attempts to use an alternating current in conjunctionwith a synchronous alternating magnetic field to counteract theelectrolysis of water. By constantly reversing the fields, bubbleformation is reduced. Unfortunately, this only provides for a minimalincrease in voltage and flow before electrolysis occurs. In addition itis much more difficult to perform. It requires precise shifts in theelectric and magnetic fields, otherwise the fluid does not flow at all.

It is therefore desirable to provide a microfluidic propulsion techniquethat requires relatively little power.

It is also desirable to provide a microfluidic propulsion technique thatutilizes a constant magnetic field.

It is also desirable to provide a microfluidic propulsion technique thatmay be used on a variety of fluids, specifically aqueous and hydrophobicsolutions and structures.

It is also desirable to provide a microfluidic propulsion technique thatdoes not induce water electrolysis.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows a schematic diagram of a magnetohydrodynamic device.

FIG. 2 shows a schematic diagram of an alternative magnetohydrodynamicdevice formed on a glass substrate.

FIG. 3 shows a cross-sectional view of the schematic diagram of FIG. 2.

FIG. 4A shows a perspective view of a magnetohydrodynamic channel.

FIG. 4B shows a perspective view of an alternative magnetohydrodynamicchannel.

FIG. 4C shows a perspective view of an alternative magnetohydrodynamicchannel.

FIG. 4D shows a perspective view of an alternative magnetohydrodynamicchannel.

FIG. 5A shows a schematic diagram of a flow pattern of a solution withina magnetohydrodynamic channel.

FIG. 5B shows a schematic diagram of an alternative flow pattern of asolution within a magnetohydrodynamic channel.

FIG. 6 shows a schematic diagram of an alternative magnetohydrodynamicdevice.

FIG. 7 shows a schematic diagram of a hole punch pattern for forming amagnetohydrodynamic device on ceramic tape.

FIG. 8 shows a schematic diagram of a magnetohydrodynamic device formedon a piece of ceramic tape.

FIG. 9 shows a cross-sectional view of the schematic diagram of themagnetohydrodynamic device of FIG. 8.

FIG. 10 shows a schematic diagram of a magnetohydrodynamic chipcomprised of ceramic tape and incorporating the ceramic tape of FIGS. 8and 9.

FIG. 11 shows a schematic diagram of a series of pieces of ceramic tapedesigned to form a magnetohydrodynamic chip having a feedback loop.

FIG. 12 shows a schematic diagram of an alternative embodiment of amagnetohydrodynamic device.

FIG. 13 shows the magnetohydrodynamic device of FIG. 12 after analytesolution has entered the main channel.

FIG. 14 shows the magnetohydrodynamic device of FIG. 12 where the deviceis pumping the analyte solution toward a reservoir.

FIG. 15 shows an alternative embodiment of a magnetohydrodynamic device.

FIG. 16 shows alternative design patterns for microfluidic channels onceramic tape.

FIG. 17 shows a series of pieces of magnetic tape that may be stacked toform the microfluidic chip shown in FIG. 18.

FIG. 18 shows a microfluidic chip capable of performing a redox assay.

FIG. 19 shows a schematic diagram of a microflidic assay structure.

FIG. 20 shows an alternative microfluidic assay structure.

FIG. 21 shows a schematic diagram of an alternative microfluidic assaystructure.

SUMMARY OF THE INVENTION

The invention disclosed here is a new method of microfluidic propulsionand a set of devices that promises to solve many of the problems thatother existing microfluidic methods suffer from. This new approach iscapable of moving small volumes of fluids through a channel, in eitherdirection, without valves. These devices may be constructed from avariety of materials and use voltages that are in the millivolt to voltrange. In addition, this invention utilizes relatively small, constantmagnetic fields that can be provided by small permanent magnets. Themethod disclosed is effective on extremely small samples, less than 100picoliters. MHD is also a readily reversible method of pumping. Thesecharacteristics make this invention especially suitable for use in LOCtechnology. It may be used for chemical analysis of very small samples,such as those common in forensics, DNA and medical testing.

In order to avoid water electrolysis, chemicals that are highlysusceptible to reduction/oxidation are added to the solution prior toits addition to the microfluidic system. These reduction/oxidation(redox) chemicals serve as “ferries” transporting electrons from theanode to the chathode. Once oxidized at the cathode, they return to theanode where they are once again reduced. This cycle is repeated manytimes. By using redox chemicals in the fluid as electron transporters,electrolysis of water is avoided. The redox species in the solution arepropelled in a direction perpendicular to both the electric and magneticfields. This movement of the redox species causes the entire solution toflow in the same direction. This constitutes a significant improvementover existing microfluidic technology.

In addition to redox species, other current carrying species may beused. Metallic nanoparticles may be added to the solution in order toaccomplish the same motion as redox compounds. The nanoparticles ferryelectrons, thereby allowing current to flow through the solution. Thisis necessary for the Lorentz forces to take effect. The movement ofnanoparticles causes the entire solution to move.

Mixing is generally difficult with extremely small volumes. Samplepreparation and assays, such as immunoassays and DNA analysis, involvecombining of reagents in very small amounts. Small samples posechallenging problems in analyzing their content, since signal of smallvolumes is generally small or requires special equipment to achieve highsensitivity. Some chemical detection methods, such as electrochemicaldetection, have a signal that depends on how fast the molecular speciesmove past the detector or a modified surface which captures the analyte.The inability to mix extremely small volumes allows diffusion topredominate and significantly reduces the accuracy of small sampleanalysis. The invention disclosed herein allows mixing of such smallsamples and can significantly improve the accuracy of small sampleanalysis. Methods of mixing extremely small samples have eludedscientists for years. In the present invention, relatively highconcentration of current carrying species, such as redox chemicals,nanoparticles, or the like are usually used. This guarantees theinhibition of water electrolysis. The current carrying species carriesthe entire current. This also rapidly and effectively mixes samples assmall as a hundred picoliters. The rapid movement of the redox speciesor nanoparticles mixes the solution.

Another advantage provided by the present invention is that a widevariety of redox chemicals may be used. It may be desirable to use MHDmicrofluidic technology to analyze chemicals that react with variousredox compounds. In such situations, a different, non-reactive redoxchemical, nanoparticle or the like may be employed instead without anyadverse effects. This ability to choose from a wide range of suitablecurrent carrying species makes the present invention more practical andmore adaptable than other microfluidic pumping methods. It is alsopossible that small scale, portable LOC devices may be exposed tovarious extreme conditions. Some conditions such as extreme pressures,heat or cold may have an adverse effect on certain redox chemicals.Again, in these situations specific redox chemicals may be employed thatbest suit a given situation.

The present invention also allows the pumping action of the MHDmicrofluidics channel to be separated from the analyte solution. Ahydrophobic solution having a current carrying species may be used topush an aqueous solution through a microfluidic channel. The twosolutions will not mix together. This provides many advantages. Manyanalytes may react with redox species or nanoparticles to form differentcompounds. This will decrease the accuracy of any measurements of theanalyte. Current carrying species may disrupt the method of detection ofthe analyte, causing false positives or false negatives. By separatingthe analyte solution from the pumping solution, the analyte remainsunaffected.

In the present invention, a small channel is formed through which thefluid flows. To avoid evaporation, the channel is enclosed on foursides. Two opposing sides consist of electrodes. It is usually desirablethat these electrodes be switched, so that each alternates between beinga cathode and an anode allowing direction of flow to reverse. A magneticfield passes through the two remaining walls, perpendicular to theelectric field created by the electrodes. A solution having a currentcarrying species is introduced to the channel. Lorentz forces affect thecurrent carrying species, propelling them in a direction perpendicularto both the electric and magnetic fields. The current carrying speciesin turn causes the solution to move through the channel.

The magnetohydrodynamic microfluidic systems disclosed herein areespecially suited for the formation of microfluidic assay structuresthat allow the performance of chemical detection assays within verysmall chips. Reduction/oxidation assays, ELISA's polynucleotidehybridizations and other immobilization assays may all be performedwithin microfluidic assay structures. These assays, because of theirelectrochemical nature and small volume, are fast, reliable, sensitiveand easily transported.

It is therefore an object of the present invention to provide aneffective method of microfluidic pumping.

It is another object of the present invention to provide a method ofusing (MHD) technology to pump microfluidic samples without inducingwater electrolysis.

It is another object of the present invention to provide a method ofrapidly and effectively mixing extremely small samples. It is anotherobject of the present invention to provide a method of pumpingmicrofluidic samples that is effective for a wide variety of samplesolutions. FEATURES OF MICROFLUIDICS Feature Electro-kinetic MechanicalCentrifugal Magnetohydrodynamic Flow and Limited Variable & Variable &Variable (slow) & non- Profile (slow) & flat non-flat non-flat flatReversible Yes Yes (valves, No Yes Direction pushed) Voltage & High(100's to For pump For Spinning 0.01 V to 1 V Power 10000's V) devicesVersatile No Yes Yes Yes Materials and Solvents Easy to Yes (device) no(moving) No (moving Yes (battery) Miniaturize, No (power) parts, valvesparts, low complexity detection)

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

The embodiments discussed herein are merely illustrative of specificmanners in which to make and use the invention and are not to beinterpreted as limiting the scope of the instant invention.

While the invention has been described with a certain degree ofparticularity, it is to be noted that many modifications may be made inthe details of the invention's construction and the arrangement of itscomponents without departing from the spirit and scope of thisdisclosure. It is understood that the invention is not limited to theembodiments set forth herein for purposes of exemplification.

MHD Lorentz forces have been known to physicists for almost 200 years.It involves 3 physical fields all perpendicular to one another. The flowor velocity field is aligned perpendicular to both the magnetic andelectric fields which are also perpendicular to one another.Manipulation of any two of these fields results in a change in the thirdone. In the present invention, an electric field and a magnetic fieldare applied to a channel both being perpendicular to the desireddirection of flow.

MHD technology requires a relatively dense current in order to induce asignificant rate of flow through the channel. At larger scales this isnot practical. Small scale, microfluidic channels however, because ofthe unique properties associated with microelectrodes in close proximityto one another, allow for relatively high current densities. These maybe combined with natural magnets. Magnetic fields on 0.4T or less may beadequate. Because natural magnets and a low amount of electricity areall that is required, MHD technology is especially well suited for LOC.

Microfluidic MHD channels may be constructed from a variety ofmaterials. Channels formed from ceramic tape and glass slides arediscussed in detail below. However, any substrates susceptible tomicrofabrication are suitable.

The current carrying species may be any chemical compound capable ofreadily acquiring and releasing one or more electrons. Those skilled inthe art of chemistry will recognize that there are a large number ofchemical compounds, generally known as redox compounds or species, thatwould serve as adequate current carrying species. Two common examples ofwell known redox compounds are ferricyanide and hydroquinone. Also,small particles, called nanoparticles, may serve as adequate currentcarrying species. Nanoparticles may be comprised of metals, carbonfibers, conductive plastics or the like. Depending on the solution towhich they are added, redox species or nanoparticles may be hydrophobic,hydrophilic or amphoteric.

FIG. 1 illustrates the principles by which the present inventionoperates. Electrodes 10 and 16 are connected to an electrical powersource 12. Electrode 16 works as a cathode while electrode 10 works asan anode. This creates an electrical field represented by directionalarrow 14. Natural magnets, not shown in this drawing, are used to applya magnetic field represented by directional arrow 18. The magnetic fieldis applied perpendicular to the electric field. Current between cathode16 and anode 10 is carried by a current carrying species 22. Currentcarrying species 22 acquires an electron from anode 10 and istransformed into the reduced form of species 24. The reduced species 24then carries the electron to cathode 16 where it discharges it andtransforms into the oxidized species 22. This process is repeated manytimes. Electric field 14 created by the redox cycling of the currentcarrying species, in conjunction with the magnetic field, induces flowof the current carrying species in the direction of directional arrow20. The flow induced within the current carrying species is transferredto the entire solution located between the electrodes.

FIGS. 2 and 3 illustrate a MHD channel formed between 2 glass slides. Toform this microchannel, an electrode is first deposited on each glassslide. An insulating material is then used to form a channel on oneslide and then the second slide is attached. The slides are positionedin such a way that the electrodes deposited each run along a wall of thechannel. Reservoirs are then placed at each end of the channel, amagnetic field is applied, and current is sent through the electrodes. Acurrent carrying species within the solution carries the current andcreates an MHD effect. In FIG. 2 MHD device 30 has an upper reservoir 32and a lower reservoir 34. Lower reservoir 34 is sealed so that it is airtight and is conected to upper reservoir 32 by pressure equalizing tube36. As solution is transferred to upper reservoir 32, a partial vacuumis created in lower reservoir 34, making it more difficult for thedevice to draw solution out of reservoir 34. Equalizing tube 36 relievesthis vacuum pressure.

Class slides 42 and 44 have electrodes 46 and 48 respectively.Electrodes 46 and 48 are on the sides of slides 42 and 44 that face eachother. This way electrodes 46 and 48 lie along 2 opposite walls of thechannel. Insulating material 38 and 40 lie between slides 42 and 44, andadhere to each of them. Insulating material 38 and 40 may be comprisedof any of a number of materials. Polydimethylsiloxane (PDMS) elastomer,polyimide and various photo resists may all be used. It is also possibleto use double sided adhesive tape for the insulating material.

FIG. 3 shows a cross sectional view of an MHD channel formed by thefollowing process:

Device Fabrication Procedure

Electrode Fabrication

1. Glass microscope slides are piranha cleaned for 30 minutes thenrinsed thoroughly with water.

2. Deposit a 100 Å chromium adhesion layer, then 6000 Å of gold on aglass microscope slide using a thermal evaporator.

3. Coat the deposited glass slides with approximately 1.5 mL of HPR-504positive photoresist.

4. Spin coat the slides for 20 seconds at 2000 rpm.

5. Bake the slides for 9 minutes at 103° C. on top of petri dishes.

6. Place the electrode photoplot film ink-side down on top of thedeposited slide and place another slide on top of the film.

7. Expose slides to UV light for 30 seconds for each half of the slide(1 minute total).

8. Develop the exposed slide for 1 minute in a 50:50 solution ofwater:developer solution.

9. Place the slides in Aqua Regia (3:1 Hcl:HNO₃) for 3-4 minutes oruntil the gold is etched away. Rinse with water.

10. Place the slides in a Chromium Etch agent for 1 minute, or until allof the chromium has been etched away. Rinse with water.

11. Rinse all remaining photoresist away with acetone and rinse withwater.

The pattern from the electrode film should now be transferred onto theglass slide.

Channel Mold Fabrication

1. A silicon wafer is piranha cleaned for 30 minutes then rinsedthoroughly with water.

2. Coat the wafer with SU-8 25 (Microchem Corp), a negative photoresist.

3. Spin coat the wafer for 30 seconds at 2000 rpm.

4. Soft bake the wafer to 5 minutes at 95° C.

5. Place the channel film ink-side down on top of the wafer and place amicroscope slide on top of the film.

6. Expose the wafer for 5 minutes.

7. Hard bake the wafer to 15 minutes at 95° C.

8. Develop the exposed wafer to 3 minutes (1 minute of agitation andsoaking for 2 minutes) in SU-8 Developer.

9. Spray with fresh developer.

10. Bake at 60° C. until dry.

The mold should be hard.

Transferring Channel to Electrode Slide

1. Mix polydimenthylsiloxane (PDMS) elastomer and curing agent (10:1 byweight) thoroughly.

2. Degas mixture for 10-30 minutes.

3. Pour mixture on top of channel mold.

4. Press electrode slide firmly against the channel mold.

5. Bake for 1-3 hours at 60° C. until cured.

6. Peel the electrode slide and the channel mold wafer apart. The PDMSshould adhere to the glass electrode slide, thereby transferring thechannel design onto the electrode slide.

7. Clear the residual PDMS out of the channels using a sharp object(i.e. razor blade).

8. Piranha clean the electrode slide with the PDMS and another electrodeslide (which has holes drilled in it for the reservoirs) for 15 minutesand rinse thoroughly with water.

9. Immediately rinse both slides with methanol and press them together(gold sides facing each other).

10. Bake at 65° C. until methanol is completely dry. The two slidesshould now be joined together.

This process creates the sandwich arrangement in FIG. 3. In theparticular embodiment described above, glass slides 52 and 54 are usedas the beginning substrate. However, any substrate susceptible to anyprocess of depositing layers of conducting material may be used.Oxidized silicon wafers and polyimide films are examples of othersuitable substrates.

The channel formed by this process may be as little as 12 microns wideand 12 microns long. However, it is also possible to form channels 12microns wide and several millimeters long. This process allows theformation of a structure having several channels in a variety ofdesigns.

Also in this embodiment, thermal evaporation is used to deposit goldelectrodes 58 and 56 onto slides 52 and 54 respectively. However, thoseskilled in the art will appreciate that there are a variety of methodsfor depositing these electrodes. Electron beam evaporation, sputteringdeposition, spin coating, molecular beam epitaxy or the like aresuitable alternatives to thermal evaporation. The preferred method ofdeposition will depend on the type of substrate used, the use to whichthe MHD device is to be put, the desired characteristics of the MHDdevice and other factors known to those skilled in the art.

Those skilled in the art will also appreciate that gold is only one ofmany suitable materials for the conducting layer. Other metals such ascopper and aluminum are suitable for use as electrodes. It may also bedesirable to use non-metallic conductors, such as carbon fibers for theelectrode layer.

Insulating layer 60 is sandwiched between slides 52 and slides 54 inorder to form channel 62. Slides 52 and 54 are positioned such thatelectrodes 58 and 56 face one another. Slides 52 and 54 are off set fromone another to facilitate attachment of conducting wire 64 and 66 thatlead to an electrical current source. In this particular embodiment,PDMS is used as the insulating layer. This layer may be as thin or asthick as desired. The only limit on the thickness of the layer is thatit must be thick enough to prevent shorting between electrodes 58 and56.

FIGS. 4A, 4B, 4C and 4D all show alternative designs for a MHD channel.In FIG. 4A, general MHD channel 70 has a basic design. Anode 76comprises 1 of 4 walls of the channel. Cathode 78 comprises the oppositewall of channel 70. Side walls 72 and 74 are comprised of an insulatingmaterial. FIG. 4A illustrates the simplest design where anode 76 andcathode 78 each comprise an entire wall of the channel.

FIG. 4B illustrates a more complex design for channel 70. In thisembodiment, wall 86 has a series of anode bands 92 running down thelength of the wall. Here there are 3 anode bands 92 but this number mayvary. Similarly, wall 84 has a series of cathode bands 80 running alongits length. Preferably there are the same number of anode bands 92 andcathode bands 80. However, this is not necessary. Side walls 88 and 90are comprised of insulating materials.

FIG. 4C shows another alternative embodiment for channel 70. In thisparticular embodiment, anode bands 104 are located on the edges of wall110 in the corners of the channel created by walls 110, 106 and 108.Similarly, cathode bands 102 are located in the corners formed betweenwall 112 and walls 106 and 108. Those skilled in the art will realizethat the different geometries found in 4A, 4B and 4C are slight and thatthe channels are substantially similar.

FIG. 4D shows an alternative embodiment that has significant differencesfrom the other illustrated embodiments. Channel 120 is specificallydesigned to alter the flow profile of the fluid within the MHD channel.Microfluidic channels impose a significant amount of drag on solutionsthat pass through them. This creates a “D” shaped flow profile. In somesituations, it may be advantageous to have a more square shaped flowprofile. Channel 120 alters the flow profile by replacing the insulatingwalls with passive equilibration conducting walls 126 and 128. Passiveequilibration conducting walls 126 and 128 contact anode wall 122 andcathode wall 124. The partial equilibration conduction caused by walls126 and 128 effect the flow pattern of the fluid within the channel 120.

The magnets used to induce the magnetic field are not shown in thesedrawings. Those skilled in the art will understand that the magnets donot need to be in actual physical contact with the channel or thesolution within the channel. It is only necessary that magnets bepositioned close to the MHD channel in order to induce a magnetic fieldin the proper orientation.

FIGS. 5A and 5B illustrate different flow patterns. FIG. 5A illustratesthe flow through general channel 70. Fluid 132 flows unevenly asindicated by flow vector arrows 134. The least amount of drag isexperienced by the portion of the fluid in the middle of the channel.This creates a cone shaped flow pattern. FIG. 5B illustrates a squareshaped flow pattern. Fluid 130 moves at an even rate as indicated byflow vector arrows 136. Passive equilibration causes this type of squareshaped flow pattern.

FIG. 6 shows a microfluidic MHD channel combined with a microcavitysensing device. Microfluidic structure 140 has a first reservoir 142, asecond reservoir 144, a microfluidic MHD channel 146 and a microcavity148. The magnet used to induce a magnetic field is not shown in theillustration. Electrodes 156 and 154 are used in conjunction with anexterior magnet to create the MHD effect. This causes solution inreservoir 142 to enter channel 146 and pass through it, eventuallyreaching reservoir 144. When the solution encounters microcavity 148,microcavity electrodes 150 and 152 may be used to detect variousanalytes. Such microcavities are described in detail in U.S. patentapplication Ser. No. 09/946,249 and U.S. patent application Ser. No.09/978,734. The microfluidic structure shown in FIG. 6 is a relativelysimple design. It may be desirable to incorporate several microcavitiesalong the wall of microfluidic MHD channels. The presence of themagnetic field also induces mixing within the microcavity. This canincrease the speed and accuracy of the detection of analytes within theanalyte solution.

In an alternative embodiment of microfluidic MHD channels, the channelsare fabricated in ceramic tape commercially available from DuPont. Themicrofluidic devices are fabricated on Green Tape™ 951 series and 851series. The designs consist of microchannels fabricated on 6 in.² piecesof substrates with gold electrodes screen printed on the sidewalls ofthe channels. The thickness of the screen print is 12 um. Theseelectrodes extend to form contact pads at the side of the chip for edgeconnectors. The gold electrodes form the electrical part of themagneto-hydrodynamic pump.

The material used in fabrication is known as Green Tape™ and is alsoknown as Low Temperature Co-fired Ceramic (LTCC). In the pre-fired statethe ceramic tape consists of alumina particles, glass frit and organicbinder. In the pre-fired state the ceramic tapes are soft, pliable andeasily machinable. Mesoscopic features ranging in size from 10 μm to 10mm can be machined using mechanical, chemical and thermal means. Thismaterial is compatible with high conductivity metals such as gold andsilver.

Green Tape™ comes in two varieties. The 951 series and the 851 seriesboth have similar compositions but the 851 is white in the pre-firedstate and the 951 is blue.

Fabrication Terminology:

Via—hold punched in ceramic tape using a punching machine.

Filled vias—vias filled with metal to form electrical interconnectsbetween layers.

Catchpads—patches of metal printed on the tape directly above filledvias to assist with the electrical interconnect. J

Registration holes—vias punched at four corners of the ceramic tape toassist in stacking of the tapes during the lamination process.

Alignment holes—vias close to the registration holes to assist withalignment during the screen print process.

Screen print—process of printing metal onto the ceramic tape.

The fabrication process for the Green Tape™ proceeds in several steps.The tape comes in a roll, which is cut into 6 in² pieces, then cured at120° C. for 30 minutes. Then registration and alignment holes arepunched along with other features necessary for the overall function ofthe device such as microchannels are vias. Each layer is fabricatedseparately. These individual layers will eventually be arranged in theproper order (stacked) to form a 3D structure. The next step afterpunching is the via fill. In this stage vias are filled with metallic,conductive ink. These help to form electrical interconnections betweenlayers. After this, screen-printing of the gold electrode ensues. Bythis stage all the microchannels have been punched on the tape and thesidewalls of the microchannel are coated with metal by pulling a vacuumthrough during the screen print (FIG. 9). After the screen print processis lamination. With the aid of the registration holes the various layersare stacked together then vacuum sealed and a hydraulic pressure of 3500psi at 80° C. is applied for 10 minutes. Then the substrate is baked at850° C. for 8 hours.

The Green Tape™ shrinks on heating. There is a 12% shrinkage in the x,yplane and 15% shrinkage in the z-axis. The shrinkage is predictable thuscan be compensated for during the design.

There are several methods available to create vias and microchannels onthe ceramic tape. These methods include milling, punching, jet vaporetching and laser machining.

A 3 dimensional channel system may be designed by stacking severallayers of the tape. The sidewalls of the channels may be coated withgold to form the electrodes for the magnetohydrodynamic pump.

FIG. 7 illustrates the use of a hole puncher to form channels andreservoirs in ceramic tape. FIG. 7 is an enlarged schematic diagram ofthe whole punching pattern used. Circular hole punch pattern 160 is usedto form reservoirs in the tape. Straight hole punch pattern 162 is usedto form a channel. By punching several holes in an overlapping manner, achannel and ring may be formed. FIG. 8 shows a top plan view of a pieceof ceramic tape 170 that has had reservoirs 172 and channel 174 punchedthrough it. Electrodes 176 are formed by screen printing conductive inkin a cross shaped pattern over the channel as shown in FIG. 8. A vacuumis applied to the opposite side of the tape. This causes the ink to rundown the sides of the channel and to separate so as to form two (2)electrodes. FIG. 9 is a cross sectional view of the same piece of tapeshown in FIG. 8 along cross section line 178. FIG. 9 shows howelectrodes 176 run along the inside of channel 174 on opposite walls.Excess conductive ink is pulled through channel 174 by the vacuum, so asto prevent channel 174 from being filled. By this method, 2microelectrodes 176 are formed within the channel.

FIG. 10 shows the same piece of ceramic tape stacked with additionalpieces of ceramic tape in order to form a microfluidic chip. Channeltape 170 shown in FIGS. 8 and 9 rest on top of support tape 180 andunderneath via tape 182. Via tape 182 has holes filled with conductivematerial positioned such that they engage electrodes 176. Resting atopvia tape 182 is top tape 184. Top tape 184 also has vias filled withconductive material. On the top side of tape 184 are catch pads 190. Thecatch pads are significantly larger than the extremely small vias 188.This is to facilitate connection to an electric current. Each catch pad190 is connected to an electric current power source, such that one actsas an anode while the other acts as a cathode. Current travels from thecatch pad through vias 188 down to electrodes 176. The current carryingspecies within the solution in channel 174 completes the circuit.

In this particular embodiment, the conductive material that theelectrodes and catch pads are comprised of an that fill the vias iseither gold or silver conductive ink. However, those skilled in the artwill understand that any conductive material that is compatible withco-fired ceramic tape will be suitable.

All 4 pieces of ceramic tape have aligning holes 186. Proper alignmentof these holes insures that the catch pads, vias and electrodes arealigned properly so that they may conduct electric current. Once theceramic tape pieces have been fabricated and aligned, they are firedtogether to form a single solid chip. Prior to firing, the tape isflexible. However, once fired the chip structure becomes rigid. Becausethe chip is very thin, they may become brittle if there are aninsufficient number of tape layers. Therefore, it is often desirable toinclude extra tape layers to strengthen the chip. FIG. 10 shows a chiphaving 4 layers. It is often more practical to form chips comprised of10 or more layers to add strength and support. These additional layersmay consist of additional support tapes, additional via tapes and/oradditional channel tapes.

FIG. 11 shows a top down schematic diagrams of alternative chipconfigurations. The embodiment shown in FIG. 11 forms a microfluidic MHDchannel having a feedback loop. Top plate 300 has reservoirs 302 thatare connected by main channel 304. Main channel electrodes 306 and 308are screen printed onto the ceramic tape such that they are connected tocatch pads 310 and 312 respectively.

Reservoirs 302 and main channel 304 are formed by the manner shown inFIG. 7. Overlapping punch holes are used to form the pattern.

Catch pads 314 and 316 are screen printed such that conductive materialconnects them to vias 318 and 320 respectfully. These vias are formedusing the same hole puncher used to form the reservoir/channel design.Four aligning holes 322 are also punched into the tape. Second tape 380has aligning holes 324 that correspond to aligning holes 322 in the toptape 300. Vias 326 and 328 are filled with conductive material andcorrespond to vias 318 and 320 respectively. Transport vias 330correspond to reservoirs 320. These vias are left hollow so that thesolution may pass through them.

Bottom plate 340 has alignment holes 332 that correspond to alignmentholes 324 and tape 380 and to holes 322 and tape 300. Conductive pattern334 is applied to bottom plate 340 such that via 326 is conductivelyconnected to feedback channel electrode 342. Similarly, conductivepattern 336 is applied to bottom tape 340 in such a way as toconductively connect via 328 to feedback channel electrode 344 when the3 tapes are stacked. Feedback channel 346 is formed by makingoverlapping hole punches as shown in FIG. 7. Once all of the holes havebeen punched in the tape and the conductive material has been applied,the 3 tapes are stacked and co-fired. As with the previous example, itis often desirable to include several additional layers of tape in orderto strengthen the final chip.

FIG. 12 shows an alternative embodiment of a microfluidic MHD channeldesigned to measure the volume of the analyte solution being analyzed.This embodiment is particularly well suited for keeping the analytesolution separate from the pumping solution. Analyte solution 208 islocated in reservoir 210. Electrodes 224 and 226, in conjunction with amagnetic field applied to the structure 200, causes analyte solution 208to flow through introduction channel 218 and into main channel 215. Mainchannel electrodes 220, 232, 234 and 222 are not active. Exit channelelectrodes 228 and 230 are active and draw pumping solution 206 from themain channel 215 through the main channel 215 toward exit channel 216 bythe vacuum caused by the pumping of the pumping solution 206 byelectrodes 228 and 230.

Once analyte solution 208 begins to enter the exit channel 216, as shownin FIG. 13, introduction channel electrodes 224 and 226 and exit channelelectrodes 228 and 230 are turned off. Main channel electrodes 220, 222,232 and 234 are turned on to cause pumping solution 206 to enter themain channel 215 from reservoir 202 and to exit the main channel 215into reservoir 214. The movement of the pumping solution 206 causes theanalyte solution 208 to travel down main channel 215 toward reservoir214, as shown in FIG. 14. Analysis of the analyte solution may occur atany point within the structure. In addition, other microfluidic channelsmay be added to main channel 215 or reservoir 214 and the analytesolution may be pumped through them.

The amount of analyte solution analyzed is determined by the distancebetween the introduction channel 218 and exit channel 216. The distancebetween these two channels multiplied by the cross-sectional are of thechannel equals the volume of analyte solution drawn into main channel215. FIG. 15 shows a microfluidic structure that operates in the samefashion as the structure shown in FIGS. 12, 13, and 14. However, instructure 400 introduction channel 404 and exit channel 406 join mainchannel 402 at the same point. This causes the portion of the analytesolution 408 that ravels to reservoir 412 to be as small as possible.Pumping solution 420 propels analyte solution 418 down main channel 402.

Pumping solutions 206 and 420 may be hydrophobic and analyte solutions208 and 418 may be hydrophilic, or vice versa. This prevents the analytesolution from mixing with the pumping solution. Those skilled in the artwill understand that there are advantages to keeping the analytesolution free of carrier species.

Other microfluidic pumping devices may be readily adapted for use inconjunction with a MHD system. The analyte solution may be introducedinto the main channel using a different type of pumping. Once theanalyte solution is within the main channel, an immiscible pumpingsolution may be used to propel the analyte solution through thestructure.

The above embodiments describe a single channel with or without a secondfeedback loop. One embodiment describes a main channel and two side(introduction and exit) channels. However, those skilled in the art willappreciate that a natural extension of these designs include a series ofmicrofluidic channels, each utilizing the same magnetic field and eachhaving independently addressable electrodes. These channels may beinterconnected so that fluids may be propelled by Lorentz forces throughmore than one or all of them. FIG. 16 shows a piece of ceramic tape 500having a number of microchannel structures. Microchannel 502 illustrateshow a microchannel may have several branches, while microchannels 504,506, 508, 510 and 512 show a variety of different microchannel patterns.

FIG. 17 show a series of pieces of ceramic tape 820, 822, 824, 826 and828. These pieces of LTCC may be stacked and fired to form a ceramicchip capable of performing redox assays. Tape 820 has a gold electrode832 stamped onto it in the form of gold ink. Gold ink circuit 834extends away from electrode 832 and is aligned with vias thatconductively connect it to a catch pad as described below. Electrode 832will serve as the bottom of assay structure 902 as shown in FIG. 18.

Chip 822 has two holes that will serve as reservoirs punched through it,as well as filled via hole 840. Hole 836 will form the bottom portion ofa sample reservoir 898 in FIG. 18. Hole 838 will form part of assaystructure 902. Via 840 is filled with conductive, metallic ink.

Central tape 824 has hole 842 that in conjunction with hole 836 formssample reservoir 898. Similarly, hole 852 in conjunction with hole 838forms assay structure 902. Holes 842 and 850 are connected by conduit848. As can be seen in FIG. 18, conduit 848 is sandwiched between tapes826 and 822 to form a magnetohydrodynamic conduit. Electrodes 844 and846 are used to form an electric field within conduit 848 such thatfluids having carrier species may be transported from sample reservoir898 to assay structure 902. Electrodes 850 and 856 are located on thesides of hole 852 and may serve as detecting electrodes within assaystructure 902. Exit conduit 854 allows a sample to be pushed out ofassay structure 902. Alternatively, conduit 854 exposes fluids withinthe microfluidic structure to the environment, thereby relieving backpressure. Filled via 870 allows electrode 832 to be connected to a catchpad.

Tape 826 consists of microfluidic conduits 858 and 868, as well asfilled vias 860, 862, 864, 866 and 872. These filled vias connectelectrodes to catch pads on the top layer piece of tape 828. Sampleconduit 858 is part of conduit 904 that allows sample to be introducedinto sample reservoir 898. Conduit 868 allows conduit 854 to be exposedto the environment, thereby relieving back pressure for allowing sampleto exit the microfluidic chip.

Top layer top 828 has hole 874 that completes conduit 904 and allowsintroduction of a sample into sample reservoir 898. Similarly, hole 876completes conduit 906. Catch pad 878 is comprised of gold ink and liesdirectly above filled via 880. Catch pad 878 is thereby conductivelylinked to electrode 832. Catch pads 894, 892, 882 and 886 are similarlyconnected to electrodes 844, 846, 850 and 856 by filled vias 860, 862,864 and 866 respectively. These catch pads allow current to beconductive to their respective electrodes. This facilitates MHDpropulsion and detection of redox cycling.

The tape layer shown in FIG. 17 is layered as shown in FIG. 18 to formmicrofluidic assay chip 830. Structure 830 is then fired so as to formthe microfluidic assay chip. Aligning holes may be added to the tapelayers, but is not shown here for clarity. Similarly, magnets areapplied to the top and bottom of chip 830 to allow MHD propulsionthrough conduit 848. The similarly is not shown for clarity, but theirpositioning would be clear to one skilled in the art.

This relatively simplistic microfluidic assay chip 830 is operated inthe following procedure. First, sample is introduced through conduit 904into sample reservoir 898. Sufficient sample is introduced such thatsome enters conduit 848 in between electrodes 844 and 846. Because thisassay is a redox assay, the sample being tested may also serve as acarrier species. An electric field is then applied to conduit 848through electrodes 844 and 846 so as to induce MHD propulsion. In thismanner, the sample will enter the assay structure 902. Conduit 906prevents the build up of back pressure. Leaving conduit 904 exposed tothe environment also relieves back pressure. Due to the relatively smalldiameters of conduits 904 and 906, the effects of evaporation aregreatly reduced.

Once the sample enters assay structure 902, redox cycling may bemeasured. Any combination of two of electrodes 850, 856 and 832 may beutilized to measure redox cycling by cyclic voltammetry or other methodsknown in the art. In this embodiment, three electrodes are presentwithin the assay structure and all three may be utilized. However, onlytwo electrodes are necessary to perform redox cycling, as is wellrecognized by a skilled artist.

Those skilled in the art will appreciate that the concentration of avariety of redox compounds may be detected by the present invention. Oneparticularly useful assay for use in the present invention is thedetection of dopamine. Measuring low concentrations of dopamine hasproven difficult because it is usually found in extracellular fluidsthat also include ascorbic acid in significantly greater concentrations.The chip described in FIG. 18 allows dopamine to constantly cyclebetween electrodes while ascorbic acid is irreversibly oxidized andcenses contributing to electric current after a few seconds.

FIG. 19 shows a diagrammatic top view of a system designed forconducting an assay within a magnetohydrodynamically driven microfluidicsystem. The scheme in FIG. 19 may be used in microfluidic systems formedwith either ceramic tape or photolithographic methods. The scheme ofFIG. 19 may be punched or etched into a single layer that is thensandwiched between upper and lower layers.

Those skilled in the art will appreciate that FIG. 19 is one of manypossible configurations for the assays disclosed herein and is thereforeessentially a schematic diagram.

The sample 612 that is being analyzed is first placed in samplereservoir 604 by means of insert port 603. This may be accomplished by avariety of methods including, but not limited to, injection by asyringe, using either a micro or macro scale pump and capillary action.If sample 612 is known to have a carrier species at a relatively highconcentration, no carrier species will need to be added to facilitateMHD propulsion. However, it may be necessary to add an appropriatecarrier species. It is also desirable that sample 612 have relativelylow concentrations of salts. Although salts may assist MHD propulsion,they lead to rapid corrosion of electrodes. It may therefore bedesirable to remove salts by precipitation or other methods known in thearts. Of course, if the microfluidic chip into which this assay isincorporated is intended to be a single use, disposable chip, corrosionof the electrodes is inconsequential.

Once sample 612 has been inserted into reservoir 604, electric currentis applied to electrodes 654 and 652 such that one is an anode and oneis a cathode, thereby creating an electric field. Permanent magnetsabove and below structure 600 (not shown) provide a magnetic field. Thiscauses sample 612 to flow through conduit 622. Those skilled in the artwill appreciate that one of the disadvantages of MHD propulsion is thatit has slow flow rates. One of the advantages of using MHD inmicrofluidic systems is that conduits through which fluids flow arerelatively short. This allows even the slow flow rate of MHD to providefor rapid analysis. Sample 612 flows through conduit 622 into assaystructure 602. Those skilled in the art will appreciate that it may bepossible to apply sample 612 directly to assay structure 602 without theneed for sample reservoir 604 or conduit 622. However, those skilled inthe art will also appreciate that whichever structure, assay structure602 or sample reservoir 604, the sample is applied to will be exposed tothe open environment and therefore subject to evaporation. Evaporationis a serious factor to consider with very small volumes. If assaystructure 602 is not exposed to the environment, the concentration willremain constant. Therefore, it is often advantageous to utilize aseparate sample reservoir. In addition, although not shown, thoseskilled in the art will appreciate that it is a relatively simple matterto add additional sample conduits to reservoir 604 that connect it toadditional assay structures such that a variety of assays may beperformed on a single sample.

Assay structure 602 has detecting electrodes 632 and 634. In thisparticular embodiment, the MHD assay chip is formed on ceramic tape.Therefore, electrodes 634 and 632 are comprised of a metallic,conductive ink that is printed onto a layer of tape. Those skilled inthe art will appreciate that if the microfluidic structure is formedusing photolithographic techniques, it may be easier and more desirableto have detecting electrodes that are formed as tubular nanobandelectrodes.

Primary analyte binding material 638 is located on either the top or thebottom (or both) of assay structure 602. Analyte binding material 638may be comprised of any of a variety of materials well known to thoseskilled in the art. In this particular embodiment, analyte bindingmaterial 638 is a primary antibody developed for an ELISA technique. Inthis particular embodiment, assay structure 600 is designed to detectthe microorganism Cryptosporidium parvum. Therefore, ABM 638 is aprimary antibody that is specific for C. parvum. It is attached to aself-assembled monolayer formed by lipids having a sulphate group ontheir hydrophilic ends. The sulphate group is covalently bound to goldink printed on the ceramic tape layer that comprises the bottom of theassay structure 602. Once sample 612 enters assay structure 602, any C.parvum present will bind to ABM 638.

Rinse reservoir 606 contains a rinsing solution 614. Rinsing solution614 may simply be Dl water. Alternatively, solution 614 may be comprisedof a series of buffers and/or salts and having a pH that optimizes theELISA being performed. Additionally, rinse solution 614 has adequateconcentration of a carrier species to facilitate MHD propulsion. Acurrent is applied to electrodes 650 and 648 such that one serves as acathode and one an anode. This in conjunction with permanent magnetsabove and below conduit 624 cause solution 614 to enter assay structure602. Conduit 624 has hydrophobic bead 630 in order to prevent prematuremixing. Those skilled in the art will appreciate that hydrophobic bead630 may not be necessary because of the relatively little mixing in amicrofluidic system. Hydrophobic bead 630 is comprised of a hydrophobicliquid, such as an oil. It serves as a plug between conduit 624 andassay structure 602.

Once sufficient time has elapsed for substantially all C. parum presentin sample 612 to bind to ABM 638, solution 614 is propelled into assaystructure 602 using MHD. Sample solution 612 and some of solution 614exits assay structure 602 through conduit 636. Conduit 636 may lead to awaste reservoir for discarded fluid. Alternatively, conduit 636 may leadto a second asssay structure.

Once assay structure 602 has been rinsed by fluid 614, solution 616 heldin reservoir 608 is then introduced to assay structure 602. In thisembodiment, fluid 616 caries a secondary ABM, in this case a secondaryantibody specific for C. parvum having an electroactive complexcovalently attached. Solution 616 is located in conduit 626 betweenhydrophobic beads 633 and 631. Beads 633 and 631 may be comprised of thesame material as beads 630 in conduit 624. 617 may be comprised of thesame solution as 616 or may be a different solution. Almost any solutionwill be suitable for 617 so long as it has an adequate concentration ofcarrier species to facilitate MHD propulsion. When a current is passedthrough electrodes 646 and 644, MHD propulsion of solution 617 isfacilitated by the electric field and magnetic field provided bypermanent magnets not shown. The presence of beads 631 and 633 preventsadditional secondary ABM from entering assay structure 602 after excesssecondary ABM has been rinsed out of assay structure 602. Furthermore,because bead 631 is downstream of electrodes 644 and 646, it is notnecessary that solution 616 have a carrier species. This reduces theamount of carrier species present in assay structure 602 and helps toreduce background noise of electrochemical measurements. Beads 633 and631 are not necessary for the present invention to function properly.However, they are generally preferred because they usually increase theaccuracy of the invention. It is possible for reservoir 608 to holdsolution 616 and for 616 to have a carrier species in it.

Once secondary ABM solution has entered assay structure 602 andsufficient time has been allowed for secondary ABM to attach to any C.parum present, assay structure 602 is again rinsed with solution 614.

This removes excess, unbound secondary ABM. The next step in the assayis to activate the electroactive complex. The method of activation willdepend upon the type of electroactive complex used. Here, theelectroactive complex is alkaline phosphatase. Therefore, activation ofthe electroactive complex consists of adding PAP to assay structure 602.

Substrate reservoir 610 holds substrate solution 620 which contains PAP.Because PAP is a redox compound, it may also double as a carrier speciesfor MHD propulsion. When a current is applied to electrodes 642 and 640,the electric field in conjunction with the magnetic field produced bymagnets above and below this structure cause substrate solution 620 toenter assay structure 602 by traveling down conduit 628. As doesconduits 624 and 626, conduit 628 has a hydrophobic bead plug 635. Aswith the other beads, this bead is not necessary but is preferred. Thoseskilled in the art will appreciate that although PAP is the substrateused for alkaline phosphatase, other electroactive complexes may requireother substrates that may or may not be redox species. Solution 620 maybe comprised of substrates or other activating compounds, such as abuffer that changes pH within the assay structure. As described above,some electroactive complexes are activated by change in pH. Activatingspecies may also be a coenzyme or cofactor. Those skilled in the artwill also appreciate that some electroactive species will not requirethe addition of a solution to be activated. In that situation, reservoir610 and conduit 628 are unnecessary.

In this particular embodiment, assay structure 602 is rinsed by solutionfrom reservoir 606 twice. Those skilled in the art will appreciate thatthis will also be accomplished by utilizing two reservoirs instead ofone.

Once the electroactive species has been activated by addition of anactivating compound or other means, current is run through electrodes632 and 634. Cyclic voltammetry or other current measuring methods maybe utilized to evaluate the elecrochemical activity of the contents ofassay structure 602. The presence of two electrodes within the assaystructure facilitates measurements of cyclic voltammetry and alsofacilitates redox cycling to amplify the electrochemical signal.

Those skilled in the art will appreciate that there will also bebackground noise. To evaluate background noise so that it may besubtracted from the signal received by electrodes 632 and 634, it may bedesirable to include a second control assay structure in a chip. Acontrol assay structure would include all of the same features ofmicrofluidic assay structure 600 except for the presence of primary ABM638. The process would be run simultaneously on the same sample. Samplefor the control may be MHD driven from reservoir 604 or may have its ownsample reservoir separate from structure 600. In addition, due to theextremely small size of assay structure 600, several such assaystructures may be placed in an array or grid pattern on a chip. Thearray would include one or more controls as well as a variety of assays.Binding materials may be antibodies specific for other microorganisms ormolecules. ABMs may also be DNA probes, protein substrates or othercompounds having specificity. In addition, the primary ABMs locatedwithin the structure that cause an analyte to immobilize mayalternatively be comprised of a binding material lacking specificity.Examples of this include polystyrene and nitrocellulose.

Assay structure 600 is designed such that electrodes 640, 642, 644, 646,648, 650, 652 and 654 are all aligned such that they may all use themagnetic fluid from a pair of large magnets located above and below theelectrodes. The magnets may be electromagnets, but permanent magnets arepreferred in order to reduce energy consumption of assay chips. Becausethe electrodes are aligned, long, narrow magnets may be used therebyboth simplifying manufacture and reducing the amounts of materialneeded. While it is preferred to utilize a structure that requires onlya single, localized magnetic field, other configurations are possible.However, utilizing multiple magnetic fields unnecessarily complicatesthis device. When an array of assays are used, it would be desirable toalign all the electrodes used for MHD propulsion either in a single longline or in a single region of the chip such that they may all use asingle magnetic field. Electrodes 632 and 634 are not associated withMHD propulsion electrodes because they are used for measuring electriccurrent and not for moving fluid within the microfluidic structure.

FIG. 20 shows a schematic diagram of an alternate embodiment of thepresent invention. Because MHD propulsion requires the presence of acarrier species, substantial additional background noise often resultsin electrochemical measurements of assays. This is because carrierspecies b their nature are capable of redox cycling. This can makeelectrochemical measurements more difficult at low analyteconcentrations. To overcome this, the present invention has describedthe techniques shown in FIGS. 12 through 15. FIG. 18 shows how thosetechniques could be applied for use in small scale assays. This improvesthe sensitivity of the assays. The structure or method is very similarto that described in FIG. 19. The significant difference between thediagrams shown in FIGS. 19 and 20 is that FIG. 20 employs “hydrophobicpumps” to inject the sample, secondary ABM solution , rinse solution andactivating solution into the assay structure.

Sample solution 663 is placed into sample solution reservoir 662. Thismay be accomplished in the same methods by which sample solution 612 isplaced in reservoir 604 of FIG. 19. However, the microfluidic assaystructure 660 of FIG. 18 requires that once sample solution 663 isplaced in reservoir 662, reservoir 662 must be sealed hermetically. Ifreservoir 662 is not sealed properly, then activation of hydrophobicpump 675 will result in a substantial portion of the sample to exitthrough entry port 661. This may be accomplished by partially fillingport 661, thereby blocking it, or by placing some sort of seal over port661.

Once reservoir 662 is no longer exposed to the outside environment,hydrophobic pump 675 is actuated. This is accomplished by applyingcurrent to electrodes 730 and 731 such that an electric field is formedbetween them. Permanent magnets (not shown) above and below conduit 684create a magnetic field in approximately the same region as conduits 684to which electrodes 730 and 731 apply an electric field. Pump reservoir676 is filled with pump fluid 677. Pump fluid 677 is hydrophobic.Hydrophobic solutions may be comprised of any hydrophobic compound solong as it is liquid at room temperature and is capable of dissolving acarrier species. Those skilled in the art will appreciate that there area variety of organic redox cycling compounds that are soluble inhydrophobic solutions. Various compounds used in cellular respirationwithin mitochondrial membranes are well suited to be these carrierspecies. When electric current is applied to electrodes 730 and 731,Lorentz forces induce MHD propulsion of the hydrophobic fluids. Thispushes the aqueous sample from sample reservoir 662 down conduit 668 andinto assay structure 664. Once sufficient sample has entered assaystructure 64, the electrical current is ceased and the hydrophobic pumpis thereby deactivated. As with the system described in FIG. 19, theassay structure 664 has a primary ABM on its bottom and attached to theunderlying piece of ceramic tape. However, in the assay described inFIG. 20, the primary ABM is a DNA probe. It is covalently bound to thehydrophobic end of a lipid that is part of a self-assembled monolayerwhich is, in turn, covalently bound on its hydrophilic end to a mentalsurface. The DNA probe is complimentary to an analyte polynucleotidestrand.

In this particular embodiment, the assay is designed to perform ahybridization assay. It is therefore desirable to include heatingelement 669 which is attached to reservoir 662. The heating elementheats the sample solution, thereby denaturing double stranded DNA. Whileit is preferred to have heating element 669, it is not necessary. It ispossible to heat and denature the sample prior to placing it withinreservoir 662.

Once sufficient time has been given for any analyte DNA to anneal to theprimary ABM probe, it is rinsed by rinse solution 714 which is stored inreservoir 670. Reservoir 670 has a hydrophobic pump attached to it bymeans of piston conduit 866. Pump reservoir 678 has a hydrophobicsolution 720 having a dissolved carrier species within it. This pumpoperates in the same way as pump 675. An electric current is applied topiston conduit 866 by electrodes 698 and 700. This works in conjunctionwith a magnetic field to cause solution 720 to move towards reservoir670 and act as a piston within conduit 866. This, in turn, pushesrinsing solution 714 down conduit 685 and into assay structure 664.Conduit 685 has a hydrophobic bead 713 to serve as a plug so thatrinsing fluid 714 does not leak into assay structure prematurely. Aswith the hydrophobic plugs in FIG. 19, this plug is not necessary but ispreferred. Once rinsing fluid 714 has flushed assay structure 664 of thesample, the electric field generated by electrode 698 and 700 is ceasedso as to stop the pumping action.

Rinsing fluid 714 may be comprised of deionized water. Because Lorentzforces are not applied to solution 714 itself, it has no need for acarrier species. This is generally preferred as it does not introduceany compounds that may produce background noise to the structure 664.

Reservoir 672 has solution 712 in which a secondary ABM, in this case asecondary probe, is dissolved. The secondary ABM probe has a carrierspecies covalently bound to it. Once the rinsing step is completed,electric current is applied to electrodes 702 and 704 to create anelectric field through piston conduit 688. Hydrophobic solution 718 hasa carrier species dissolved within it and is kept in reservoir 680. Anelectric field is applied that works conjointly with a magnet to pushsolution 718 down piston conduit 714 to work as a piston and pushsolution 712 out of reservoir 672, down conduit 686 and into assaystructure 664. Solution 712 may also be comprised of buffers and/orsalts and/or chelating agents and other compounds known to those skilledin the art to enhance the hybridization process. Once sufficientquantities of solution 712 have entered assay structure 664, theelectric current applied to electrodes 702 and 704 is cased and pumpingaction stops. As with the other plugs described herein, hydrophobic bead713 is not necessary but is preferred. This microfluidic assay structurealso shows that it is not necessary to have a second bead in the conduitthat introduces the secondary ABM.

After sufficient time has been allowed for the secondary ABM to bind toany analyte DNA present, assay structure 664 is again rinsed by rinsesolution 714. As with the initial rinse, addition of rinse solution 714to assay structure 664 is facilitated by applying current to electrodes698 and 700, thereby actuating a hydrophobic pump. The electric currentis ended after assay structure 664 is sufficiently rinsed.

Once excess secondary ABM is removed from assay structure 664, it may benecessary to add an activating agent and activating solution 710. Inthis particular embodiment, the electroactive species is again alkalinephosphatase. Therefore, PAP must be added to serve as a redox cyclingcompound to facilitate detection. Activating solution 710, stored inreservoir 674, is pushed by hydrophobic pump solution 716, stored inreservoir 682, when an electrical field is generated by electrodes 706and 708. Hydrophobic solution 716 moves down pump conduit 690, therebypushing solution 710 down conduit 687 and into assay structure 664. Aswith other conduits, hydrophobic bead 711 prevents premature mixing ofthe activating solutions. Once the activating solution is added,electrodes 724 and 726 are used to measure currents generated by anyalkaline phosphatase present.

As with the assay described in FIG. 19, the electroactive complexutilized in the assay described in FIG. 20 may not require an activatingagent.

FIGS. 19 and 20 disclose only two of a wide variety of immobilizationassays. They are intended to illustrate methods by which immobilizationassays may be incorporated into microfluidic systems to quickly,efficiently and easily perform assays on a wide variety of samples andanalytes. They may be used in conjunction with each other and may becombined in a variety of ways. One such combination is shown in FIG. 21.

FIG. 21 shows a method of performing two assays in sequence using amicrofluidic structure. Reservoirs 816 are actuated to deposit solutionsinto assay structures 806 and 809 by hydrophobic pumps 812 in the samemethods described above. Assay structure 806 is designed to perform anELISA substantially the same as that described above. Assay structure806 is designed to perform an ELISA substantially the same as thatdescribed in FIG. 19. Those skilled in the art will appreciate that themodifications are slight. Assay structure 806 has heating element 810attached to it. After the ELISA is completed, heating element 810 isactuated in order to heat shock immobilized microorganisms. The heatshock results in release of heat shock mRNAs. The solution in assaystructure 806, now including heat shock mRNAs is pumped down conduit 807and into assay structure 809. Assay structure 809 is substantially thesame as that described in FIG. 18. It is used to detect the presence ofany heat shock mRNAs. This combination of assays allows a microfluidicchip to rapidly and accurately detect not only the presence but also theviability of the microorganism for which it is testing. As with theassay structures disclosed in FIGS. 17 and 18, the structure disclosedin FIG. 19 may also be used to form an array of similar structures on achip. Even relatively large assay structures, such as the one disclosedin FIG. 21, take up only a few millimeters on a chip.

Microfluidic assay systems disclosed in FIGS. 17 through 21 are intendedto illustrate their use when comprised of ceramic tape. Those skilled inthe art will appreciate that similar structures are readily formed byphotolithographic methods. In addition, reservoirs and assay structuresare all depicted as being circular in shape. Those skilled in the artwill appreciate that they may take on any of a variety of forms.Similarly, conduits are all shown to be substantially straight. Thoseskilled in the art will appreciate that these conduits may be curved.The assay structures have also been shown to exist two dimensionally.Those skilled in the art will appreciate that both ceramic tape chipsand photolithographic chips are comprised of several layers. Thereservoirs and assay structures may be comprised of one or severallayers. In addition, it is not necessary that an assay system in amicrofluidic chip have a planar design. In many situations, it may bedesirable for the microfluidic assay system to utilize reservoirs frommany different layers within a chip. Similarly, conduits may readily bedesigned to penetrate several layers. Those skilled in the art willappreciate that the microfluidic assay designs schematically depicted inFIGS. 17 through 21 may be readily adapted to a large variety of threedimensional geometries.

For clarity, the magnets used to create the magnetic field are not shownin FIGS. 17 through 21. However, those skilled in the art willappreciate that a magnetic field is readily applied to the structures byplacing a pair of magnets about them. One magnet would go above thestructure and one below, both being laid at the location correspondingto points where MHD electrodes are placed along conduits.

Whereas, the present invention has been described in relation to thedrawings attached hereto, it should be understood that other and furthermodifications, apart from those shown or suggested herein, may be madewithin the spirit and scope of this invention.

1. A method for constructing a magnetohydrodynamic microfluidic device,comprising the steps of: fabricating at least one channel layer using atleast one layer of ceramic; forming at least two electrodes in at leastone channel; inserting said at least one channel layer between at leasttwo sandwiching layers; and forming electrical connectors such thatcurrent or voltage can be applied to said electrodes.
 2. The method forconstructing a magnetohydrodynamic microfluidic device according toclaim 1 including an additional step of laminating said at least onechannel layer between said sandwiching layers.
 3. The method forconstructing a magnetohydrodynamic microfluidic device according toclaim 2 wherein said step of laminating is accomplished by pressure,thermal or adhesive means.
 4. The method for constructing amagnetohydrodynamic microfluidic device according to claim 1 whereinsaid at least one layer of ceramic is a low-temperature, co-firedceramic material or a high-temperature ceramic material.
 5. The methodfor constructing a magnetohydrodynamic microfluidic device according toclaim 1 wherein at least one of said sandwiching layers is ceramic,glass or a polymer.
 6. The method for constructing a magnetohydrodynamicmicrofluidic device according to claim 5 wherein said ceramic of said atleast one of said sandwiching layers is a low-temperature, co-firedceramic material or a high-temperature ceramic material.
 7. The methodfor constructing a magnetohydrodynamic microfluidic device according toclaim 1 wherein said step of forming electrical connectors isaccomplished by fabricating circuits onto said at least one channellayer such that said circuits allow current or voltage to be applied tosaid electrodes.
 8. The method for constructing a magnetohydrodynamicmicrofluidic device according to claim 7 wherein said step offabricating circuits onto said at least one channel layer is by screenprinting, thermal deposition, sputter deposition, electron-beamdeposition, laser ablation, photolithography, electro-plating orelectroless-plating processes.
 9. The method for constructing amagnetohydrodynamic microfluidic device according to claim 1 whereinsaid step of forming electrical connectors is accomplished byfabricating circuits onto at least one of said sandwiching layers suchthat said circuits allow current or voltage to be applied to saidelectrodes.
 10. The method for constructing a magnetohydrodynamicmicrofluidic device according to claim 10 wherein said step offabricating circuits onto said at least one of said sandwiching layersis by screen printing, thermal deposition, sputter deposition,electron-beam deposition, laser ablation, photolithography,electro-plating or electroless-plating processes.
 11. The method forconstructing a magnetohydrodynamic microfluidic device according toclaim 1 wherein said electrodes are fabricated by screen printing,thermal deposition, sputter deposition, electron-beam deposition, laserablation, photolithography, electro-plating or electroless-platingprocesses.
 12. The method for constructing a magnetohydrodynamicmicrofluidic device according to claim 1 wherein said electrodes arefabricated into spatially separate brands, disks, rectangles, or othergeometric shapes to which electrical current or voltage can beindividually applied.
 13. The method for constructing amagnetohydrodynamic Microfluidic device according to claim 1 whereinsaid electrodes are fabricated using continuous conductive material ornanoparticulate material.
 14. The method for constructing amagnetohydrodynamic microfluidic device according to claim 1 whereinsaid electrodes are formed using pull-through methods.
 15. The methodfor constructing a magnetohydrodynamic microfluidic device according toclaim 1 wherein said electrodes are formed by coating both sides of saidat least one channel layer and using pull-through methods.
 16. Themethod for constructing a magnetohydrodynamic microfluidic deviceaccording to claim 1 including the additional step of fabricatingpassive equilibration conducting walls within said at least one channel.17. The method for constructing a magnetohydrodynamic microfluidicdevice according to claim 16 wherein said passive equilibrationconducting walls are constructed of continuous conductive material ornanoparticulate material.
 18. The method for constructing amagnetohydrodynamic microfluidic device according to claim 16 whereinsaid passive equilibration conducting walls are fabricated by screenprinting, thermal deposition, sputter deposition, electron-beamdeposition, laser ablation, photolithography, electro-plating orelectroless-plating processes.
 19. The method for constructing amagnetohydrodynamic microfluidic device according to claim 16 whereinsaid passive equilibration conducting walls are fabricated intospatially separated bands, disks, rectangles, or other geometric shapes.20. A magnetohydrodynamic microfluidic device, comprising: at least onechannel layer using at least one layer of ceramic; at least twoelectrodes in at least one channel; and wherein said at least onechannel layer is between at least two sandwiching layers to form aunitary device.
 21. The magnetohydrodynamic microfluidic device of claim20 wherein said at least one channel layer is laminated between saidsandwiching layers.
 22. The magnetohydrodynamic microfluidic device ofclaim 21 wherein said lamination of said at least one channel layerbetween said sandwiching layers is accomplished by pressure, thermal oradhesive means.
 23. The magnetohydrodynamic microfluidic device of claim20 wherein at least one of said sandwiching layers is ceramic, glass, ora polymer.
 24. The magnetohydrodynamic microfluidic device of claim 23wherein said ceramic of said at least one of said sandwiching layers isa low-temperature, co-fired ceramic material or a high-temperatureceramic material.
 25. The magnetohydrodynamic microfluidic device ofclaim 20 wherein said at least one layer of ceramic is alow-temperature, co-fired ceramic material or a high-temperature ceramicmaterial.
 26. The magnetohydrodynamic microfluidic device of claim 20further including electrical connectors wherein said electricalconnectors allow current or voltage to be applied to said electrodes.27. The magnetohydrodynamic microfluidic device of claim 26 wherein saidelectrical connectors are formed by fabricating circuits onto said atleast one channel layer such that said circuits allow current or voltageto be applied to said electrodes.
 28. The magnetohydrodynamicmicrofluidic device of claim 27 wherein said fabrication of saidcircuits onto said at least one channel layer is by screen printing,thermal deposition, sputter deposition, electron-beam deposition, laserablation, photolithography, electro-plating or electroless-platingprocesses.
 29. The magnetohydrodynamic microfluidic device of claim 26wherein said electrical connectors are formed by fabricating circuitsonto at least one of said sandwiching layers such that said circuitsallow current or voltage to be applied to said electrodes.
 30. Themagnetohydrodynamic microfluidic device of claim 29 wherein saidfabrication of said circuits onto said at least one of said sandwichinglayers is by screen printing, thermal deposition, sputter deposition,electron-beam deposition, laser ablation, photolithography,electro-plating or electroless-plating processes.
 31. Themagnetohydrodynamic microfluidic device of claim 20 wherein saidelectrodes are in spatially separate bands, disks, rectangles, or othergeometric shapes to which electrical current or voltage can beindividually applied.
 32. The magnetohydrodynamic microfluidic device ofclaim 20 wherein said electrodes are fabricated by screen printing,thermal deposition, sputter deposition, electron-beam deposition, laserablation, photolithography, electro-plating or electroless-platingprocesses.
 33. The magnetrohydrodynamic microfluidic device of claim 20wherein said electrodes are constructed of continuous conductivematerial or nanoparticulate material.
 34. The magnetohydrodynamicmicrofluidic device of claim 20 wherein said electrodes are formed usingpull-through methods.
 35. The magnetohydrodynamic microfluidic device ofclaim 20 wherein said electrodes are formed by coating both sides ofsaid at least one channel layer and using pull-through methods.
 36. Themagnetohydrodynamic microfluidic device of claim 20 further includingpassive equilibration conducting walls within said at least one channel.37. The magnetohydrodynamic microfluidic device of claim 36 wherein saidpassive equilibration conducting walls are constructed of continuousconductive or nanoparticulate materials.
 38. The magnetohydrodynamicmicrofluidic device of claim 36 wherein said passive equilibrationconducting walls are in spatially separate bands, disks, rectangles, orother geometric shapes.
 39. The magnetohydrodynamic microfluidic deviceof claim 36 wherein said passive equilibration conducting walls arefabricated by screen printing, thermal deposition, sputter deposition,electron-beam deposition, laser ablation, photolithography,electro-plating or electroless-plating processes.